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BackgroundIdentification of gene variants plays an important role in research on and diagnosis of genetic diseases. A combination of enrichment of targeted genes and next-generation sequencing (targeted DNA-HiSeq) results in both high efficiency and low cost for targeted sequencing of genes of interest.Methodology/Principal FindingsTo identify mutations associated with genetic diseases, we designed an array-based gene chip to capture all of the exons of 193 genes involved in 103 genetic diseases. To evaluate this technology, we selected 7 samples from seven patients with six different genetic diseases resulting from six disease-causing genes and 100 samples from normal human adults as controls. The data obtained showed that on average, 99.14% of 3,382 exons with more than 30-fold coverage were successfully detected using Targeted DNA-HiSeq technology, and we found six known variants in four disease-causing genes and two novel mutations in two other disease-causing genes (the STS gene for XLI and the FBN1 gene for MFS) as well as one exon deletion mutation in the DMD gene. These results were confirmed in their entirety using either the Sanger sequencing method or real-time PCR.Conclusions/SignificanceTargeted DNA-HiSeq combines next-generation sequencing with the capture of sequences from a relevant subset of high-interest genes. This method was tested by capturing sequences from a DNA library through hybridization to oligonucleotide probes specific for genetic disorder-related genes and was found to show high selectivity, improve the detection of mutations, enabling the discovery of novel variants, and provide additional indel data. Thus, targeted DNA-HiSeq can be used to analyze the gene variant profiles of monogenic diseases with high sensitivity, fidelity, throughput and speed.

The functions of miR‐182‐5p in the pathogenesis of diabetic nephropathy (DN) remain largely unclear. Here, we studied the roles and relationship between miR‐182‐5p and CD2AP in the development of DN. We used real‐time polymerase chain reaction (PCR) to compare miR‐182‐5p expression between DN and control groups, while computational analysis and luciferase assays were used to confirm CD2AP as a miR‐182‐5p target. Western blot and real‐time PCR were then used to measure the messenger RNA (mRNA) and protein expression of CD2AP in the presence of miR‐182‐5p. The results showed that miR‐182‐5p was highly expressed in cells isolated from people with DN. In addition, the luciferase activity of cells transfected with wild‐type/mutant CD2AP confirmed CD2AP as a direct target of miR‐182‐5p. The expression levels of CD2AP mRNA and protein were much lower in the DN group compared with that in the normal group. In addition, the expression levels of CD2AP mRNA and protein were evidently increased by a miR‐182‐5p inhibitor, but notably downregulated by miR‐182‐5p mimics or CD2AP small interfering RNA (siRNA). Furthermore, miR‐182‐5p and CD2Ap siRNA significantly reduced the survival rate and viability of transfected cells, while the miR‐182‐5p inhibitor exhibited an opposite effect. These findings indicated the presence of a negative regulatory relationship between miR‐182‐5p and CD2AP in podocytes cells and suggested that the overexpression of miR‐182‐5p contributes to the pathogenesis of DN.

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Sarcomatoid carcinoma (SC) is a rare primary malignant tumor in which both carcinomatous and sarcomatous elements occur. It can occur in many different organs and anatomical locations, such as the skin, thyroid gland, bone, urinary tract, breast, pancreas, liver and other areas. Of them, pulmonary sarcomatoid carcinoma (PSC) is a rare malignant cancer composed of sarcoma and sarcoma-like tumors with spindle or giant cell features. Here a case of a 75-year-old Chinese man with a six-month history of cough and hemoptysis is reported. Chest X-ray showed a tumor shadow in the left lung field. Chest computed tomography (CT) scan showed a lobulated mass in his left hilum and even the left pulmonary artery. Pleomorphic interstitial cells were found by bronchoscopic brushing. To establish a definitive diagnosis for PSC, a left pneumonectomy was performed. The pathological stage was IIB (pT2N1M0) based on the tumor node metastasis (TNM) staging system. The tumor's pathology, histology, immunohistochemistry and treatment methods are discussed.